title: Endogenous IgG affects the cell biology of RPE cells and involves the TLR4 pathway.
abstract: OBJECTIVE
  RPE is a key component of the blood-ocular barrier (BOB) and is equipped with immunological molecules such as toll-like receptors (TLRs) and complement receptors, which together orchestrate the innate and adaptive immunity of the eye. Immunoglobulin G (IgG) in the aqueous humor and vitreous body has traditionally been thought to be derived from serum via transcytosis across the BOB. Our previous work validated production of endogenous IgG by RPE cells locally. However, the function and role of this IgG in the intraocular immunity is poorly understood.
  
  
  METHODS
  After confirming IgG production in a human RPE cell line (ARPE-19) with immunofluorescence, in situ hybridization, and RT-PCR, we further investigated the function of endogenous IgG in RPE biology with MTS, flow cytometry, and cell invasion analysis after downregulation of IgG by siRNA. Involvement of the TLR4 pathway was also studied using Western blot, ELISA and confocal microscopy.
  
  
  RESULTS
  Endogenous IgG is crucial for support of proliferation, mitosis, migration, and inhibition of apoptosis of RPE. Moreover, production of endogenous IgG by RPE is regulated by the TLR4 pathway in a concentration- and duration-dependent manner, and IgG affects the activation of the TLR4 pathway in a synergistic manner. Activation of the FcγR I pathway and production of IL-10 could be induced by IgG derived from RPE.
  
  
  CONCLUSIONS
  These data suggest that endogenous IgG may be a molecule that is essential for the physiological function of RPE, and suggest IgG is important for regulating intraocular immune responses under physiologic and pathologic conditions.
authors.affiliation: Department of Pathology, Weifang Medical University, Weifang, China.
authors.forename: Na
authors.initials: N
authors.lastname: Niu
authors.forename: Jie
authors.initials: J
authors.lastname: Zhang
authors.forename: Michael A
authors.initials: MA
authors.lastname: McNutt
mesh_terms: D056704:Adaptive Immunity; D017209:Apoptosis; D015153:Blotting, Western; D002460:Cell Line; D004797:Enzyme-Linked Immunosorbent Assay; D005786:Gene Expression Regulation; D006801:Humans; D007074:Immunoglobulin G / Q000096:biosynthesis* / Q000276:immunology; D017403:In Situ Hybridization; D018613:Microscopy, Confocal; D034741:RNA, Small Interfering / Q000235:genetics*; D055213:Retinal Pigment Epithelium / Q000166:cytology / Q000276:immunology / Q000378:metabolism*; D020133:Reverse Transcriptase Polymerase Chain Reaction; D051197:Toll-Like Receptor 4 / Q000096:biosynthesis / Q000235:genetics*
chemical_list: D007074:Immunoglobulin G; D034741:RNA, Small Interfering; D051197:Toll-Like Receptor 4
keywords: FcγR I; RPE; TLR4; endogenous IgG; immunity homeostasis